【作者】 符水； 王丽军； 田建袅；

【Author】 Shui Fu;Lijun Wang;Jianniao Tian;School of Chemistry and Pharmaceutical Science of Guangxi Normal University;

【机构】 广西师范大学化学与药学学院；

【摘要】 随着核酸信号放大技术的发展,近来已报道了各种酶辅助放大耦合NMM插入G-四链体增强荧光信号的方法检测DNA^[1]、腺苷^[2]等。本文基于ExoⅢ辅助循环放大,实现了HIV-DNA的灵敏检测,原理如Fig.1（A）所示,HP探针包含靶识别区域Ⅰ、Ⅱ、G-四链体序列Ⅲ。无靶时,ExoⅢ不切割,NMM无法结合封闭的G-四链体,背景值（F₀）低。靶存在时,HP特异性识别靶,在ExoⅢ作用下DNA循环,从而释放大量自由的信号探针,最终结合NMM增强荧光（F）。实验结果如Fig.1（B）、（C）所示,在0.8¹5 nmol/L范围内,靶浓度C与F-F₀呈良好的线性关系,线性回归方程为:F-F₀=6.59372 C（nmol/L）-5.40457（R²=0.994）,LOD低至310 pmol/L。该方法简单,灵敏,选择性好,为HIV早期临床诊断提供了广阔的发展空间。更多还原

【Abstract】 With the development of nucleic acid amplification techniques,there had been reported many means for detection of DNA^[1],adenosine^[2] based on enzyme-assisted amplification coupling with NMM inserted into G-quadruplex enhancing fluorescent signal.In this work,we realized the sensitive detection of HIV-DNA based on ExoⅢ-assisted cycling.Schematic illustration is shown in Fig.1（A）.In the absence of target,enzymolysis was restrained,so NMM couldn’t bind with passivating G-quadruplex,resulting in low background（F₀）.In the presence of target,HP-Target was digested by ExoⅢ,then cycling produced a large amount of G-quadruplex-NMM complex（F）.Within 0.8¹5 nmol/L,target concentration and F-F₀ had good linear relationship.The linear regressive equation was F- F₀ = 6.59372 C- 5.40457（R²=0.994） with a LOD of 310 pmol/L.Our strategy with simplicity,satisfactory sensitivity and selectivity will offer a potential in HIV early clinical diagnosis.更多还原